The CASP9 Knockout HGC-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the CASP9 gene has been disrupted in the HGC-27 human gastric carcinoma cell background. This polyclonal pool comprises a heterogeneous mixture of HGC-27 derivatives, each carrying distinct loss-of-function alterations at the targeted CASP9 locus, providing a robust model for studying the consequences of CASP9 ablation at the population level.
The HGC-27 cell line is a well-characterized human gastric carcinoma model isolated from a metastatic lymph node of a gastric cancer patient. These cells maintain an epithelial morphology and are widely utilized in studies of metastatic gastric cancer, offering a physiologically relevant context for interrogating tumor cell intrinsic mechanisms, including apoptosis signaling and therapeutic resistance.
CASP9 is the apical initiator caspase of the intrinsic apoptotic pathway. Upon mitochondrial outer membrane permeabilization, cytochrome c (CYCS) is released and associates with APAF1 to form the apoptosome, where CASP9 undergoes autocatalytic activation. Active CASP9 then cleaves executioner caspases CASP3 and CASP7, which target substrates such as PARP1 and CAD to execute apoptosis. The activity of CASP9 is tightly controlled by upstream BCL2 family members, inhibitors including XIAP and Survivin (BIRC5), and the antagonist SMAC/DIABLO. Moreover, CASP9-mediated apoptosis integrates with p53/TP53 signaling and the PI3K/AKT pathway, where AKT1 phosphorylation directly suppresses CASP9 function.
Disruption of CASP9 in HGC-27 gastric carcinoma cells cripples the intrinsic apoptotic machinery, thereby generating a valuable loss-of-function model for exploring apoptosis resistance and chemotherapeutic response. Given the metastatic origin of HGC-27, this knockout system is particularly suited for investigating the role of apoptotic signaling in gastric cancer progression and the development of acquired drug resistance. The polyclonal nature of the product captures a broad spectrum of editing events, enabling researchers to assess phenotypic heterogeneity in CASP9-disrupted cells and mimic the diverse loss-of-function scenarios that may arise in tumors.
This CASP9 knockout polyclonal population is optimally designed for a variety of experimental applications, including dissection of intrinsic apoptotic signaling, validation of CRISPR/Cas9 editing, and high-throughput chemosensitivity screens. Researchers can monitor apoptosis via Western blot detection of cleaved CASP9, CASP3, and PARP1, flow cytometric Annexin V/PI staining, and caspase-3/7 activity assays. Viability assessment by MTT assay and RT-qPCR analysis of CASP9 mRNA levels further characterize the knockout. Additionally, cytochrome c release immunofluorescence and drug sensitivity profiling with cisplatin and 5-fluorouracil can elucidate the impact of CASP9 loss on therapeutic response in gastric cancer. For additional technical information, please contact Ascent Research.
